pXL_dSSN2_EF1a-tNGFR-2A-Puro
(Plasmid #201112)

• Purpose: Dual barcoding cassettes for SSN-seq; Barcode Set 2
• Depositing Lab: Pawel Mazur
• Publication: Lu et al Nat Biomed Eng. 2023 Aug 31. doi: 10.1038/s41551-023-01085-3.

Backbone

• Vector backbone: lentiCRISPR v2
• Total vector size (bp): 11123
• Modifications to backbone: The sgRNA cassette was replaced with the dual-SSN cassettes. The EFS-NS promoter was replaced with EF1a. tNGFR-P2A was introduced upstream of Puro.
• Vector type: Lentiviral
• Selectable markers: Puromycin ; truncated NGFR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: SSN.guide
• Insert Size (bp): 182
• Promoter: Human U6

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: LKO.1 5'

Gene/Insert 2

• Gene/Insert name: SSN.F30
• Insert Size (bp): 143
• Promoter: Mouse U6

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: mU6-F

Gene/Insert 3

• Gene/Insert name: tNGFR
• Species: H. sapiens (human)
• Promoter: EF1a

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: EF-1a Forward
• 3′ sequencing primer: WPRE-R

Resource Information

• Supplemental Documents:
  • pXL_dSSN2_EF1a-tNGFR-2A-Puro.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry